Genome Biology (Mar 2022)

Prime-seq, efficient and powerful bulk RNA sequencing

  • Aleksandar Janjic,
  • Lucas E. Wange,
  • Johannes W. Bagnoli,
  • Johanna Geuder,
  • Phong Nguyen,
  • Daniel Richter,
  • Beate Vieth,
  • Binje Vick,
  • Irmela Jeremias,
  • Christoph Ziegenhain,
  • Ines Hellmann,
  • Wolfgang Enard

DOI
https://doi.org/10.1186/s13059-022-02660-8
Journal volume & issue
Vol. 23, no. 1
pp. 1 – 27

Abstract

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Abstract Cost-efficient library generation by early barcoding has been central in propelling single-cell RNA sequencing. Here, we optimize and validate prime-seq, an early barcoding bulk RNA-seq method. We show that it performs equivalently to TruSeq, a standard bulk RNA-seq method, but is fourfold more cost-efficient due to almost 50-fold cheaper library costs. We also validate a direct RNA isolation step, show that intronic reads are derived from RNA, and compare cost-efficiencies of available protocols. We conclude that prime-seq is currently one of the best options to set up an early barcoding bulk RNA-seq protocol from which many labs would profit.

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